Hydrocarbons are readily oxidized by bacteria utilizing the AlkB gene and thus the actions of AlkB represent one of nature's best defenses against the environmental effects of oil spills. The mechanism by why AlkB works is not well understood. Nor is it well understood why AlkB, which is thought to be structurally similar to enzymes that catalyze the conversion of single C-C bonds to double C=C bonds, catalyzes the addition of an OH group to an alkane instead. The goal of this project is to determine the three-dimensional structure of AlkB, using X-ray crystallography, to develop a model of the relationship between structure and function in this important metalloenzyme. To accomplish this goal, mechanistic work using diagnostic substrates, and spectroscopic characterizations, will also be carried out. PUBLIC HEALTH RELEVANCE: This project focuses on understanding how the enzyme that catalyzes the first step in the transformation of oil to carbon dioxide works. It will also explore how this enzyme, which is structurally similar to enzymes important in human lipid metabolism, functions in comparison with that important class of enzymes as well.